De novo assembly and characterization of a highly degenerated ZW sex chromosome in the fish Megaleporinus macrocephalus

Carolina Heloisa de Souza Borges, Ricardo Utsunomia, Alessandro M Varani, Marcela Uliano da Silva, Lieschen Valeria Guerra, Arno Juliano Butzge, John Fredy Gomez Agudelo, Shisley Manso, Milena Vieira Freitas, Raquel Belini Ariede, Vito Antonio Mastrochirico-Filho, Carolina Penaloza, Agustin Barria, Fabio Porto-Foresti, Fausto Foresti, Ricardo Shohei Hattori, Yann Guiguen, Ross D. Houston, Diogo Teruo Hashimoto
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Abstract

Background: Megaleporinus macrocephalus (piaucu) is a Neotropical fish within Characoidei that presents a well-established heteromorphic ZZ/ZW sex-determination system and thus, constitutes a good model for studying W and Z chromosomes in fishes. We used PacBio reads and Hi-C to assemble a chromosome-level reference genome for M. macrocephalus. We generated family segregation information to construct a genetic map, pool-seq of males and females to characterize its sex system, and RNA-seq to highlight candidate genes of M. macrocephalus sex determination. Results: M. macrocephalus reference genome is 1,282,030,339 bp in length and has a contig and scaffold N50 of 5.0 Mb and 45.03 Mb, respectively. Based on patterns of recombination suppression, coverage, Fst, and sex-specific SNPs, three major regions were distinguished in the sex chromosome: W-specific (highly differentiated), Z-specific (in degeneration), and PAR. The sex chromosome gene repertoire was composed of genes from the TGF-beta family (amhr2, bmp7) and Wnt/beta-catenin pathway (wnt4, wnt7a), and some of them were differentially expressed. Conclusions: The chromosome-level genome of piaucu exhibits high quality, establishing a valuable resource for advancing research within the group. Our discoveries offer insights into the evolutionary dynamics of Z and W sex chromosomes in fish, emphasizing ongoing degenerative processes and indicating complex interactions between Z and W sequences in specific genomic regions. Notably, amhr2 and bmp7 are potential candidate genes for sex determination in M. macrocephalus.
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从头组装并鉴定巨头鱼高度退化的 ZW 性染色体
背景:巨头鱼(Megaleporinus macrocephalus,piaucu)是一种新热带鱼类,属于Characoidei科,具有完善的异形ZZ/ZW性别决定系统,因此是研究鱼类W和Z染色体的良好模型。我们利用 PacBio reads 和 Hi-C 为大头鲂组装了染色体水平的参考基因组。我们生成了家系分离信息以构建遗传图谱,对雌雄个体进行pool-seq以描述其性别系统的特征,并通过RNA-seq突显大口鲈性别决定的候选基因:结果:大头蝠参考基因组长度为1,282,030,339 bp,等位基因和支架N50分别为5.0 Mb和45.03 Mb。根据重组抑制模式、覆盖率、Fst 和性别特异性 SNPs,性染色体可分为三个主要区域:W特异性(高度分化)、Z特异性(退化)和PAR。性染色体基因库由 TGF-beta 家族(amhr2、bmp7)和 Wnt/beta-catenin 通路(wnt4、wnt7a)的基因组成,其中一些基因具有差异表达:piaucu 染色体级基因组具有很高的质量,为推进该群体的研究提供了宝贵的资源。我们的发现为鱼类 Z 和 W 性染色体的进化动态提供了见解,强调了正在进行的退化过程,并显示了特定基因组区域中 Z 和 W 序列之间复杂的相互作用。值得注意的是,amhr2和bmp7是巨头鱼性别决定的潜在候选基因。
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